Use of an enzyme-linked immunosorbent assay (ELISA) for screening of hybridoma antibodies against cell surface antigens.
A micro-ELISA for screening of antibodies from hybridoma cultures against surface antigens of human melanoma is described. The technique employs alkaline phosphatase-conjugated protein A and target cells attached to poly-L-lysine-coated microtiter plates. The micro-ELISA is equally sensitive as the radioimmunoassay. Mild glutaraldehyde treatment of cells did not lead to an appreciable loss of antigen activity. The fixed cells can be stored at 4 degrees C for at least 6 weeks. It is concluded that the ELISA is superior to the radioimmunoassay in the following aspects: (1) exclusion of radioactive hazards, (2) speed of performance, and (3) lower costs.
D000818:Animals; D000906:Antibodies; D000937:Antigen-Antibody Reactions; D000954:Antigens, Surface; D002470:Cell Survival; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005976:Glutaral / Q000494:pharmacology; D006801:Humans; D006822:Hybrid Cells / Q000276:immunology*; D008545:Melanoma / Q000276:immunology; D051379:Mice; D011863:Radioimmunoassay; D013997:Time Factors
D000906:Antibodies; D000954:Antigens, Surface; D005976:Glutaral